TLR4信号促进前列腺癌PC3细胞VEGF/IL-8分泌及相关信号机制  

TLR4 signaling promotes secretion of VEGF/IL-8 in prostate cancer PC3 cells and related mechanism

作　　者：郭栋[1] 刘秋燕[1] 

机构地区：[1]第二军医大学免疫学研究所,医学免疫学国家重点实验室,上海200433

出　　处：《第二军医大学学报》2010年第7期697-701,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30771984;30972688)~~

摘　　要：目的研究TLR4在人前列腺癌PC3细胞中的表达意义及相关胞内信号机制。方法利用TLR4特异性配体脂多糖(LPS)刺激人前列腺癌PC3细胞,分别于LPS刺激0、2、6、12、24、48h后收集细胞和上清,通过RT-PCR和蛋白质印迹方法检测TLR4在基因和蛋白水平的表达变化,通过RT-PCR方法检测胞内TGF-β、VEGF、IL-8、COX-2和MMP3的mRNA表达变化和ELISA方法检测上清VEGF、IL-8蛋白的表达变化;为了进一步研究相关信号通路,分别采用MAPK和NF-κB信号通路抑制剂预处理PC3细胞,然后利用同等浓度的LPS刺激,分别于4h和24h后收集上清,通过RT-PCR和ELISA方法重复上述细胞因子的检测。结果在LPS刺激后,人前列腺癌PC3细胞的TLR4功能性表达上调,引起胞内TGF-β、VEGF、IL-8、COX-2和MMP3的mRNA表达升高和上清中VEGF、IL-8的分泌增多(P<0.05);进一步研究表明胞内p38MAPK和NF-κB信号通路参与了此过程。结论 TLR4信号通过p38MAPK和NF-κB信号通路促进VEGF和IL-8的分泌。Objective To study TLR4 expression in human prostate cancer PC3 cells and the related intracellular signaling mechanisms.Methods Human prostate cancer PC3 cells were stimulated with TLR4-specific ligand lipopolysaccharide（LPS）,then the cells and supernatants were collected 0,2,6,12,24,and 48 hours after LPS stimulation.TLR4 mRNA and protein expression was examined by reverse transcription-PCR and Western blotting analysis,respectively.The mRNA expression of TGF-β,VEGF,IL-8,COX-2,and MMP3 was also measured by reverse transcription-PCR,and the levels of VEGF,IL-8 in the supernatants were examined by ELISA.To further study the related signaling pathway,MAPK and NF-κB signaling pathways were blocked by specific inhibitors in PC3 cells before LPS stimulation;the cells were collected after 4 hours and the supernatants were collected after 24 hours;and the above mentioned factors were examined by reverse transcription-PCR and ELISA again.Results TLR4 expression was up-regulated by LPS stimulation in human prostate cancer PC3 cells,which significantly increased mRNA expression of TGF-β,VEGF,IL-8,COX-2,and MMP3 and secretion of VEGF and IL-8 in the supernatants（P0.05）;further study showed that p38 MAPK and NF-κB signal pathways were involved in the process.Conclusion TLR4 signaling promotes VEGF and IL-8 secretion through p38 MAPK and NF-κB signal pathways.

关 键 词：前列腺肿瘤 TOLL样受体4 血管内皮生长因子受体 白细胞介素8 信号转导 

分 类 号：R737.25[医药卫生—肿瘤]